No abstract

No abstract

As environmental and genetic components contribute to the PCOS expression, we compared levels of endocrine disruptors, steroid hormones, cytokines, and metabolic parameters in twenty healthy, nine normal-weight PCOS women, and ten obese PCOS women. Steroid hormones, bisphenols (BPA, BPS, BPF, BPAF) and parabens (methyl-, ethyl-, propyl-, butyl-, benzyl-parabens) were measured by liquid chromatography-tandem mass spectrometry. Differences between the groups were assessed using the Mann-Whitney U test. Spearman correlation coefficients were calculated for the individual parameters relationship. Significantly higher levels of BPA, anti-Müllerain hormone, lutropine, lutropine/folitropine ratio, testosterone, androstenedione, 7β-OH-epiandrosterone, and cytokines (IL-6, VEGF, PDGF-bb), were found in normal-weight PCOS women compared to controls. Between normal-weight and obese PCOS women, there were no differences in hormonal, but in metabolic parameters. Obese PCOS women had significantly higher insulin resistance, fatty-liver index, triglycerides, cytokines (IL-2, IL-13, IFN-γ). In healthy, but not in PCOS, women, there was a positive correlation of BPA with testosterone, SHBG with lutropine, and folitropine, while testosterone negatively correlated with SHBG. In obese women with PCOS, insulin resistance negatively correlated with SHBG and estradiol. No differences were observed in the paraben exposure. Levels of BPA were higher in PCOS women, indicating its role in the etiology. Obesity significantly worsens the symptoms.

Steroid profiling helps various pathologies to be rapidly diagnosed. Results from analyses investigating steroidogenic pathways may be used as a tool for uncovering pathology causations and proposals of new therapeutic approaches. The purpose of this study was to address still underutilized application of the advanced GC-MS/MS platform for the multicomponent quantification of endogenous steroids. We developed and validated a GC-MS/MS method for the quantification of 58 unconjugated steroids and 42 polar conjugates of steroids (after hydrolysis) in human blood. The present method was validated not only for blood of men and non-pregnant women but also for blood of pregnant women and for mixed umbilical cord blood. The spectrum of analytes includes common hormones operating via nuclear receptors as well as other bioactive substances like immunomodulatory and neuroactive steroids. Our present results are comparable with those from our previously published GC-MS method as well as the results of others. The present method was extended for corticoids and 17α-hydroxylated 5α/β-reduced pregnanes, which are useful for the investigation of alternative "backdoor" pathway. When comparing the analytical characteristics of the present and previous method, the first exhibit by far higher selectivity, and generally higher sensitivity and better precision particularly for 17α-hydroxysteroids. Key wordsSteroid metabolome • Human blood • Gas chromatographytandem mass spectrometry • Backdoor pathway • Pregnancy • Mixed umbilical cord blood Methods SamplesSerum samples from non-pregnant subjects were collected from the employees of the Institute of Endocrinology, Prague, Czech Republic and their relatives, as well as from patients of the Institute of Endocrinology. Serum samples from pregnant women and umbilical cord serum at birth were obtained from patients

Endocrine disruptors (EDs) are known to have harmful effects on the human endocrine system; special effort is actually given to the exposure during pregnancy. Humans are usually exposed to a mixture of EDs, which may potentiate or antagonize each other, and the combined effect may be difficult to estimate. The main phthalate monoesters monoethyl-, mono-n-butyl-, monoisobutyl-, monobenzyl-, mono-(2-ethylhexyl)-, mono-(2-ethyl-5-hydroxyhexyl)- and mono-(2-ethyl-5-oxohexyl) phthalate were determined in 18 maternal (37th week of pregnancy) and cord plasma samples using liquid chromatography-tandem mass spectrometry. Previously determined levels of selected bisphenols, parabens and steroids were also considered in this study. In cord blood, there were significantly higher mono-n-butyl phthalate levels than in maternal blood (p=0.043). The results of multiple regression models showed that maternal plasma phthalates were negatively associated with cord plasma androstenedione, testosterone and dehydroepiandrosterone and positively associated with estradiol and estriol. For estriol, a cumulative association was also observed for Σbisphenols. To the best of our knowledge, this is the first pilot study evaluating the effect of prenatal exposure by multiple EDs on newborn steroidogenesis. Our results confirmed phthalate accumulation in the fetal area and disruption of fetal steroidogenesis. This preliminary study highlights the negative impacts of in utero EDs exposure on fetal steroidogenesis.

After menopause, when estrogen levels decrease, there is room for the activity of anthropogenic substances with estrogenic properties – endocrine disruptors (EDs) – that can interfere with bone remodeling and changes in calcium-phosphate metabolism. Selected unconjugated EDs of the bisphenol group – BPA, BPS, BPF, BPAF, and the paraben family – methyl-, ethyl-, propyl-, butyl-, and benzyl-parabens – were measured by high performance liquid chromatography-tandem mass spectrometry in the plasma of 24 postmenopausal women. Parameters of calcium-phosphate metabolism and bone mineral density were assessed. Osteoporosis was classified in 14 women, and 10 women were put into the control group. The impact of EDs on calcium-phosphate metabolism was evaluated by multiple linear regressions. In women with osteoporosis, concentrations of BPA ranged from the lower limit of quantification (LLOQ) – 104 pg/ml and methyl paraben (MP) from LLOQ – 1120 pg/ml. The alternative bisphenols BPS, BPF and BPAF were all under the LLOQ. Except for MP, no further parabens were detected in the majority of samples. The multiple linear regression model found a positive association of BPA (β=0.07, p<0.05) on calcium (Ca) concentrations. Furthermore, MP (β=-0.232, p<0.05) was negatively associated with C-terminal telopeptide. These preliminary results suggest that these EDs may have effects on calcium-phosphate metabolism.

Dental composite materials often contain monomers with bisphenol A (BPA) structure in their molecules, e.g. bisphenol-A glycidyl dimethacrylate (Bis-GMA). In this study, it was examined whether dental restorative composites could be a low-dose source of BPA or alternative bisphenols, which are known to have endocrine-disrupting effects. Bis-GMA-containing composites Charisma Classic (CC) and Filtek Ultimate Universal Restorative (FU) and “BPA-free” Charisma Diamond (CD) and Admira Fusion (AF) were examined. Specimens (diameter 6 mm, height 2 mm, n=5) were light-cured from one side for 20 s and stored at 37 °C in methanol which was periodically changed over 130 days to determine the kinetics of BPA release. BPA concentrations were measured using a dansyl chloride derivatization method with liquid chromatography – tandem mass spectrometry detection. The amounts of BPA were expressed in nanograms per gram of composite (ng/g). BPA release from Bis-GMA-containing CC and FU was significantly higher compared to “BPA-free” CD and AF. The highest 1-day release was detected with FU (15.4±0.8 ng/g), followed by CC (9.1±1.1 ng/g), AF (2.1±1.3 ng/g), and CD (1.6±0.8 ng/g), and the release gradually decreased over the examined period. Detected values were several orders of magnitude below the tolerable daily intake (4 µg/kg body weight/day). Alternative bisphenols were not detected. BPA was released even from “BPA-free” composites, although in significantly lower amounts than from Bis-GMA-containing composites. Despite incubation in methanol, detected amounts of BPA were substantially lower than current limits suggesting that dental composites should not pose a health risk if adequately polymerized.

Polycarbonates are polymers of bisphenol A (BPA), a well-known endocrine disruptor. This study evaluated the release of BPA from polycarbonate crowns that were (1) milled from Temp Premium Flexible (ZPF, Zirkonzahn, Italy) or Tizian Blank Polycarbonate (TBP, Schütz Dental, Germany), or (2) 3D-printed (Makrolon 2805, Covestro, Germany). Commercial prefabricated polycarbonate crowns (3M, USA) and milled poly(methyl methacrylate) (PMMA) crowns (Temp Basic, Zirkonzahn, Italy) were included for comparison. The crowns were stored at 37 °C in artificial saliva (AS) or methanol, which represented the worst-case scenario of BPA release. Extracts were collected after 1 day, 1 week, 1 month and 3 months. BPA concentrations were measured using liquid chromatography-tandem mass spectrometry. The amounts of released BPA were expressed in micrograms per gram of material (μg/g). After 1 day, the highest amounts of BPA were measured from milled polycarbonates, TBP (methanol: 32.2 ± 3.8 mg/g, AS: 7.1 ± 0.9 mg/g) and ZPF (methanol 22.8 ± 7.7 mg/g, AS: 0.3 ± 0.03 mg/g), followed by 3D-printed crowns (methanol: 11.1 ± 2.3 mg/g, AS: 0.1 ± 0.1 mg/g) and prefabricated crowns (methanol: 8.0 ± 1.6 mg/g, AS: 0.07 ± 0.02 mg/g). Between 1 week and 3 months, the average daily release of BPA in methanol and AS decreased below 2 mg/g and 0.6 mg/g, respectively. No BPA was released from PMMA in AS, and the cumulative amount released in methanol was 0.2 ± 0.06 mg/g. In conclusion, polycarbonates could be a relevant source of BPA, but the current tolerable daily intake of BPA (4 mg/kg body weight) should not be exceeded.